This SBIR Phase I project will determine the feasibility of applying polypeptide multilayer nanofilm technology to synthetic biomatrix development and commercialization. These nanofilms are a type of surface coating for cell and tissue culture. Immediate in vitro uses of synthetic biomatrix materials include many areas of basic medical research, diagnostic testing, high-throughput drug screening, cell phenotype selection, biologics production, and development of various therapeutic cell technologies. The synthetic biomatrix market has an estimated annual value of close to $1 billion worldwide. Any part of a tissue that is not a cell is called the extracellular matrix (ECM). In the living body the ECM surrounds cells and influences cell shape, proliferation, activation, and survival. Scientific research has revealed numerous molecular signals whereby the ECM affects cell behavior. Research has also shown that the surface on which cells are grown plays a key role in determining cell behavior in vitro and in optimizing culture conditions for a particular cell type. A variety of methods are used to modify surfaces for in vitro cell culture. In one approach, one or more proteins from mammalian ECM, for example collagen, are deposited by non-specific adsorption. Such coatings offer relatively advanced functionality, but ECM proteins are expensive to purify, they must be refrigerated, they are prepared from an animal source, and they have a limited ability on their own to control cell behavior in vitro. More complex products such as MatrigelTM are useful for some purposes, but they are poorly defined with regard to molecular composition and unsuited for many purpose. The function of a synthetic biomatrix is to mimic some features of the ECM without the use of animal products and in a way that is highly defined with regard to molecular composition. The polypeptide multilayer nanofilms of the propose research offer significant advantages over existing coatings for in vitro cell culture. These nanofilms are entirely synthetic, they can mimic crucial features of ECM proteins, and they can be tailored to a particular cell type and culture objective, important for control of some aspects of cell behavior in vitro. Moreover, these nanofilms are likely to be shelf-stable at room temperature for extended periods and cost competitive with existing products in the marketplace. The proposed research has two main parts: optimization of a reliable process of polypeptide nanofilm fabrication in multi-well plates, and determination of the effect of different physical properties and biofunctionalization of these nanofilms on the proliferation, attachment, and morphology of various cell types. Feasibility of applying polypeptide multilayer nanofilm technology to synthetic biomatrix development and commercialization will have been demonstrated if polypeptide multilayer nanofilms are shown to enable reliable control over cell attachment and proliferation relative to coating products currently on the market.Project Narrative In vitro culture of mammalian cells has become indispensable for medical research, diagnostics, drug screening, large-scale production of therapeutic proteins, and other purposes touching on healthcare;interest in development of cell culture methods continues to grow rapidly. Polypeptide multilayer nanofilms are promising for the development of synthetic biomatrix materials, or artificial extracellular matrices. Such materials offer comparable or superior control over cell properties in vitro, extended shelf-life at room temperature, and cost-competitive manufacture, and they can advance knowledge of biological processes in health and disease and promote the development of new protein and cellular therapeutics.